Density dependence of daily activity in three ungulate species

Daily activity in herbivores reflects a balance between finding food and safety. The safety‐in‐numbers theory predicts that living in higher population densities increases safety, which should affect this balance. High‐density populations are thus expected to show a more even distribution of activity—that is, spread—and higher activity levels across the day. We tested these predictions for three ungulate species; red deer (Cervus elaphus), roe deer (Capreolus capreolus), and wild boar (Sus scrofa). We used camera traps to measure the level and spread of activity across ten forest sites at the Veluwe, the Netherlands, that widely range in ungulate density. Food availability and hunting levels were included as covariates. Daily activity was more evenly distributed when population density was higher for all three species. Both deer species showed relatively more feeding activity in broad daylight and wild boar during dusk. Activity level increased with population density only for wild boar. Food availability and hunting showed no correlation with activity patterns. These findings indicate that ungulate activity is to some degree density dependent. However, while these patterns might result from larger populations feeling safer as the safety‐in‐numbers theory states, we cannot rule out that they are the outcome of greater intraspecific competition for food, forcing animals to forage during suboptimal times of the day. Overall, this study demonstrates that wild ungulates adjust their activity spread and level based on their population size.     

Deciphering the products of evolution at the species level: the need for an integrative taxonomy

Progress in molecular techniques together with the incorporation of phylogenetic analyses of DNA into taxonomy have caused an increase in the number of species’ discoveries in groups with morphological characters that are difficult to study or in those containing polytypic species. But some emerged criticisms plead for a taxonomic conservatism grounded either on the requirement of providing evidences of morphological distinctiveness or reproductive barriers to erect new species names. In a case study of taxonomic research on Neotropical frogs, we combine several lines of evidence (morphological characters, prezygotic reproductive isolation and phylogenetic analyses of mitochondrial DNA) to test the status of 15 nominal species and to assess the degree of agreement of the different lines of evidence. Our study reveals that morphology alone is not sufficient to uncover all species, as there is no other single line of evidence independently. Full congruence between lines of evidence is restricted to only four out of the 15 species. Five species show congruence of two lines of evidence, whereas the remaining six are supported by only one. The use of divergence in morphological characters seems to be the most conservative approach to delineate species boundaries because it does not allow the identification of some sibling reciprocally monophyletic species differing in their advertisement calls. The separate analysis of differences in advertisement calls (evidence of reproductive isolation) or of phylogenetic data alone also shows limitations, because they do not support some morphological species. Our study shows that only an integrative approach combining all sources of evidence provides the necessary feedback to evaluate the taxonomic status of existing species and to detect putative new ones. Furthermore, the application of integrative taxonomy enables the identification of hypotheses about the existence of species that will probably be rejected or changed, and those that can be expected to persist.     

Measurement of fine root tissue density: a comparison of three methods reveals the potential of root dry matter content

Aims

Root tissue density (RTD, the ratio of root dry mass to root volume) is a fundamental trait in comparative root ecology, being increasingly used as an indicator of plant species’ resource use strategy. However, the lack of standardized method to measure this trait makes comparisons tricky. This study aims to compare three methods commonly used for determining fine RTD and to test whether root dry matter content (RDMC, the ratio between root dry mass and root fresh mass) could be used as a surrogate of fine root tissue density.

Methods

RTD of 163 fine root samples was determined using (i) Archimedes’ method, (ii) image analysis (WinRHIZO software), and (iii) using the root dry matter content as a proxy. Root samples belonged to different herbaceous species grown in different conditions.

Results

RTD measured with Archimedes’ method was positively correlated with RTD estimated with image analysis and with RDMC. However we demonstrated that RTD measured with Archimedes’ method was better predicted by RDMC (R²?=?0.90) than by RTD measured with image analysis (R²?=?0.56). The performance and limitations of each method were discussed.

Conclusion

RDMC is a quick, cheap and relatively easy measurable root attribute; we thus recommended its measurement as a proxy of fine root tissue density.     

High-speed atomic force microscopy: Imaging and force spectroscopy

Atomic force microscopy (AFM) is the type of scanning probe microscopy that is probably best adapted for imaging biological samples in physiological conditions with submolecular lateral and vertical resolution. In addition, AFM is a method of choice to study the mechanical unfolding of proteins or for cellular force spectroscopy. In spite of 28 years of successful use in biological sciences, AFM is far from enjoying the same popularity as electron and fluorescence microscopy. The advent of high-speed atomic force microscopy (HS-AFM), about 10 years ago, has provided unprecedented insights into the dynamics of membrane proteins and molecular machines from the single-molecule to the cellular level. HS-AFM imaging at nanometer-resolution and sub-second frame rate may open novel research fields depicting dynamic events at the single bio-molecule level. As such, HS-AFM is complementary to other structural and cellular biology techniques, and hopefully will gain acceptance from researchers from various fields. In this review we describe some of the most recent reports of dynamic bio-molecular imaging by HS-AFM, as well as the advent of high-speed force spectroscopy (HS-FS) for single protein unfolding.     

Plant–arthropod interactions in gymnosperm leaves from the Early Permian of Patagonia,Argentina

Evidence of plant–arthropod interactions in three orders of gymnosperms is documented in at least five localities from Lower Permian strata of the Río Genoa Formation (Patagonia, Argentina). Arthropod damages were identified in Glossopteris, Gangamopteris, Cordaites and Gingkoites leaves, corresponding to four interactive categories: margin feeding, hole feeding, surface feeding, and oviposition. In addition, a Permian insect trace not previously recorded is described on Glossopteris wilsonii leaves. The diversity of arthropod damage is consistent with the diverse flora of the Río Genoa Formation. In turn, the patterns of arthropod and plant–arthropod interaction provide constraints on the ecology and paleoclimate of Western Gondwana during the Early Permian.     

Evaluation of Macroscopic and Microscopic Components of Partial Resistance to Leaf Rust in Durum Wheat

Leaf rust, caused by the fungus Puccinia triticina, is considered one of the most important foliar diseases in durum wheat. Hypersensitive resistance (HR) may be rapidly overcome by the pathogen when resistant cultivars are grown on a large acreage or following changes in virulence in the pathogen population. Prolonging the durability of the resistance requires uses of other types of resistance such as partial resistance (PR). In this study, six durum wheat lines provided by the International Center for Corn and Wheat Improvement (CIMMYT) with a high level of PR to leaf rust were studied in monocyclic tests in a growth chamber. Inoculations were performed on both primary and fifth leaves using the Spanish race DGB/BN. UV fluorescence microscopy was employed to determine microscopic components of the resistance, such as the number of early aborted infection units not associated with plant cell necrosis (EA?) and relative colony size (RCS) of the established infection units. Macroscopic components of PR such as latency period, infection frequency and uredinium size were measured as well. All six resistant lines were characterized by a higher EA? and smaller RCS respect to the susceptible control ‘Don Rafael’. Line 3 showed the highest level of PR. It had 22% of EA? compared with 4% in the susceptible control, and the smallest RCS (17% respect to RCS of ‘Don Rafael’) at adult plant stage. Both EA? and RCS had a high heritability (more than 97%) and the correlation with macroscopic parameters (latency period and uredinium size) was also high (significant at 0.001 level). Hence, PR to leaf rust in these durum wheat genotypes has been revealed at microscopic level (higher EA? and smaller RCS).     

RNA isolation from loquat and other recalcitrant woody plants with high quality and yield

RNA isolation is difficult in plants that contain large amounts of polysaccharides and polyphenol compounds. To date, no commercial kit has been developed for the isolation of high-quality RNA from tissues with these characteristics, especially for fruit. The common protocols for RNA isolation are tedious and usually result in poor yields when applied to recalcitrant plant tissues. Here an efficient RNA isolation protocol based on cetyltrimethylammonium bromide (CTAB) and two successive precipitations with 10 M lithium chloride (LiCl) was developed specifically for loquat fruits, but it was proved to work efficiently in other tissues of loquat and woody plants. The RNA isolated by this improved protocol was not only of high purity and integrity (A₂₆₀/A₂₈₀ ratios ranged from 1.90 to 2.04 and A₂₆₀/A₂₃₀ ratios were > 2.0) but also of high yield (up to 720 μg on average [coefficient of variation = 21%] total RNA per gram fresh tissue). The protocol was tested on loquat fruit (different stages of development, postharvest, ripening, and bruising), leaf, root, flower, stem, and bud; quince fruit and root; grapevine cells in liquid culture; and rose petals. The RNA obtained with this method is amenable to enzymatic treatments and can be efficiently applied for research on gene characterization, expression, and function.